Molecular Discovery Research, GlaxoSmithKline Pharmaceuticals, New Frontiers Science Park, Harlow, Essex, CM19 5AD, UK.
Transgenic Res. 2011 Apr;20(2):429-33. doi: 10.1007/s11248-010-9413-6. Epub 2010 Jun 12.
The increasing use of ES cell lines from strains other than 129, in particular C57BL/6, has greatly reduced the time taken to generate gene knockouts on a defined genetic background. Generally, C57BL/6 ES cell lines transmit less efficiently through the germline than 129 lines; consequently the burden on animal husbandry at this stage is increased. Genotyping sperm from chimaeric males may be used to identify mice which are transmitting the manipulated allele, however it requires that the mice be culled and the sperm used for IVF. Here we describe a quick and reliable method for genotyping copulatory plugs. Males which produce a positive result can then be naturally mated. Thus far we have observed a perfect correlation between copulatory plug genotype and germline transmission, accompanied by considerable savings in mouse numbers and resources.
越来越多的人使用除 129 以外的品系的胚胎干细胞系，特别是 C57BL/6，这大大缩短了在特定遗传背景下生成基因敲除的时间。通常情况下，C57BL/6 胚胎干细胞系通过种系传递的效率低于 129 系；因此，在这个阶段对畜牧业的负担增加了。对嵌合体雄性的精子进行基因分型可以用来鉴定传递被操作等位基因的小鼠，但这需要对小鼠进行安乐死，并使用精子进行 IVF。在这里，我们描述了一种快速可靠的交配栓基因分型方法。产生阳性结果的雄性可以自然交配。到目前为止，我们已经观察到交配栓基因型与种系传递之间存在完美的相关性，同时还节省了大量的小鼠数量和资源。
